Zhang P, Wang H, Cao H, Xu X, Sun T
Department of Ophthalmology, Shanghai General Hospital, Shanghai JiaoTong University; Shanghai Key Laboratory of Fundus Disease; Shanghai Engineering Center for Visual Science and Photomedicine, Shanghai 200080, China.
Zhonghua Yan Ke Za Zhi. 2017 Mar 11;53(3):207-211. doi: 10.3760/cma.j.issn.0412-4081.2017.03.012.
To explore the inhibitory effect of insulin-like growth factor binding protein-related protein 1 (IGFBP-rP1), a novel anti-angiogenic factor, on retinal angiogenesis and its underlying molecular mechanisms. Experimental study. C57BL/6J mice were classified into three groups: control group (24), oxygen-induced retinopathy (OIR) non-intervention group (24) and OIR intervention group (72). The OIR mouse model was established using improved Smith's methods ( 96). Twelve-day-old mice in the OIR intervention group were randomly assigned into three groups receiving intravitreal injection of recombinant mouse IGFBP-rP1 (50 μg/L, 100 μg/L and 200 μg/L, respectively). Five days later, the proliferative neovascular responses were estimated by quantifying the new vessel area relative to the total retinal area in flattening retinas stained by high molecular FITC-Dextran and counting the number of neovascular cell nuclei breaking through the internal limiting membrane (ILM) in cross-sections. Retinal phosphor-ERK1/2 (p-ERK1/2), ERK1/2 and vascular endothelial growth factor (VEGF) protein expression was assessed by Western blot. In the fluorescence angiograms, irregular neovascularization and fluorescence leakage were observed in the OIR model. In the OIR non-intervention group, the expression of p-ERK1/2 and VEGF was significantly up-regulated in comparison with the control group (100.068, 0.000. 6.526, 0.003). The area ratios of new retinal vessels and the number of neovascular cell nuclei in mice receiving intravitreal injection of recombinant mouse IGFBP-rP1 both decreased significantly (1920, 0.000. 852.387, 0.000), following the down-regulation of retinal p-ERK1/2 protein expression (859.587, 0.000) and VEGF protein expression (24.301, 0.000) in a dose-dependent manner (0.05). There was no significant difference in ERK1/2 protein expression (0.05). IGFBP-rP1 inhibits retinal angiogenesis by blocking ERK signaling pathway and down-regulating VEGF expression. This highlights the potential importance of IGFBP-rP1 serving as a target of gene therapy for retinal neovascularization. .
探讨新型抗血管生成因子胰岛素样生长因子结合蛋白相关蛋白1（IGFBP-rP1）对视网膜血管生成的抑制作用及其潜在分子机制。实验研究。将C57BL/6J小鼠分为三组：对照组（24只）、氧诱导视网膜病变（OIR）非干预组（24只）和OIR干预组（72只）。采用改良的史密斯方法建立OIR小鼠模型（96）。将OIR干预组12日龄小鼠随机分为三组，分别接受玻璃体腔内注射重组小鼠IGFBP-rP1（分别为50μg/L、100μg/L和200μg/L）。五天后，通过在高分子FITC-葡聚糖染色的扁平视网膜中量化新生血管面积相对于总视网膜面积，并在横切面上计数突破内界膜（ILM）的新生血管细胞核数量，来评估增殖性新生血管反应。通过蛋白质印迹法评估视网膜磷酸化ERK1/2（p-ERK1/2）、ERK1/2和血管内皮生长因子（VEGF）蛋白表达。在荧光血管造影中，在OIR模型中观察到不规则新生血管形成和荧光渗漏。在OIR非干预组中，与对照组相比，p-ERK1/2和VEGF的表达显著上调（100.068，0.000；6.526，0.003）。接受玻璃体腔内注射重组小鼠IGFBP-rP1的小鼠视网膜新生血管面积比和新生血管细胞核数量均显著降低（1920，0.000；852.387，0.000），视网膜p-ERK1/2蛋白表达（859.587，0.000）和VEGF蛋白表达（24.301，0.000）呈剂量依赖性下调（P<0.05）。ERK1/2蛋白表达无显著差异（P>0.05）。IGFBP-rP1通过阻断ERK信号通路和下调VEGF表达来抑制视网膜血管生成。这突出了IGFBP-rP1作为视网膜新生血管化基因治疗靶点的潜在重要性。
